Identification of two fungi strains and their control effects to cereal cyst nematode
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摘要:目的
生防菌株的种类鉴定及对禾谷孢囊线虫Heterodera avenae的防治效果。
方法以形态学特征和rDNA-ITS序列比对及系统发育树分析相结合的方法鉴定生防菌的种类，测定菌株A1和B1发酵液对孢囊、卵和2龄幼虫的活性及对小麦孢囊线虫病的盆栽防治效果。
结果菌株A1和B1分别鉴定为寄生曲霉Aspergillus parasiticus和长枝木霉Trichoderma longilbrachiatum。处理14 d后，A1和B1的菌丝对线虫孢囊的寄生率可达80.00%以上。处理12 d后，菌株A1和B1的发酵液原液对卵孵化的抑制率分别为63.98%和67.56%，处理72 h后，对2龄幼虫的校正死亡率分别为68.14%和92.98%。盆栽试验发现，菌株A1和B1的3%菌肥培养物处理90 d后，孢囊减少率分别为31.71%和36.04%，小麦根长分别增加28.82%和59.62%，株高分别增加20.38%和21.43%。
结论菌株A1和B1对禾谷孢囊线虫具有防治作用，是2株极具开发潜力的生防菌株。
Abstract:ObjectiveTo identify the species of biocontrol strains and analyze their control efficiency to cereal cyst nematodes (Heterodera avenae).
MethodBiocontrol fungus A1 and B1 were identified according to morphological characters, rDNA-ITS sequence alignment and analysis of phylogenetic tree. The control effects of the fermentation broths of strain A1 and B1 against cysts, eggs and the second-stage juveniles (J2) and pot control effects against cereal cyst nematodes were determined.
ResultStrain A1 and B1 were identified as Aspergillus parasiticus and Trichoderm longilbrachiatum respectively. The mycelium parasitic rates of A1 and B1 on nematode cysts were over 80.00% at 14 days after treatment. At 12 days after treatment, the inhibition rates of the fermentation broths of A1 and B1 on egg hatching were 63.98 % and 67.56% respectively. The corrected mortality rates of J2 at 72 hours after treatment were 68.14% and 92.98 % respectively. At 90 days after treatment with 3% fungus fertilizer, pot experiment results showed that cyst reduction rates of A1 and B1 were 31.71% and 36.04%, the root lengths of wheat plants increased by 28.82% and 59.62%, and the plant heights increased by 20.38% and 21.43%, respectively.
ConclusionStrain A1 and B1 have control effects on cereal cyst nematodes and are worthy of development and utilization .
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图 1 菌株A1(a~c)、B1(d~f)的培养性状及形态特征
a、d: 菌落正面，b、e: 菌落背面，c、f: 分生孢子梗；标尺=10 μm
Figure 1. Cultural characteristics and morphologies of strain A1(a−c) and B1(d−f)
a and d are face views of colonies, b and e are reverse views of colonies, c and f are conidiophore; Bar=10 μm
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图 3 不同浸染时间A1、B1分生孢子悬浮液对禾谷孢囊线虫的寄生率
Figure 3. Parasitic rates of A1 and B1 conidia suspensions to cereal cyst nematode at different time of infection
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图 4 真菌发酵液对禾谷孢囊线虫卵孵化的影响
图柱上方不同小写字母表示差异显著(P<0.05，LSD法)
Figure 4. Effects of fungal fermentation broth on egg hatching of cereal cyst nematode
Different lowercase letters indicate significant difference(P<0.05, LSD method)
表 1 真菌发酵液对禾谷孢囊线虫2龄幼虫的致死作用
Table 1 Lethal effects of fungal fermentation broth on the second-stage juveniles of cereal cyst nematode
菌株
Straint/h 最终稀释倍数
Final dilution ratio线虫死亡数量1)
Number of dead nematode死亡率/%
Mortality rate校正死亡率1)/%
Corrected mortality rateA1 24 1× 79.33±2.33a 39.67 39.36±1.17a 5× 40.67±9.68b 20.33 19.93±4.87b 10× 34.67±1.67b 17.33 16.92±0.84b CK 1.00±0.58c 0.50 48 1× 107.33±4.91a 53.67 51.52±2.46a 5× 49.33±8.57b 24.67 22.43±4.30b 10× 37.33±1.86b 18.67 16.42±0.93b CK 4.67±0.88c 2.30 72 1× 138.00±5.51a 69.00 68.14±2.83a 5× 53.00±11.68b 26.50 24.46±0.06b 10× 39.67±1.45b 19.83 17.61±0.75b CK 5.33±0.88c 2.70 B1 24 1× 95.00±3.21a 47.50 47.24±1.62a 5× 87.00±9.17a 43.50 43.22±4.61a 10× 70.67±7.69a 35.33 35.01±3.86 CK 1.00±0.58b 0.50 48 1× 125.00±4.51a 62.50 60.38±2.26a 5× 105.67±12.99ab 52.83 50.69±6.52a 10× 79.00±8.14b 39.50 37.31±4.09b CK 4.67±0.88c 2.30 72 1× 186.33±4.33a 93.17 92.98±2.23a 5× 105.67±12.57b 52.83 51.52±6.46b 10× 84.67±4.33b 42.33 40.73±2.23b CK 5.33±0.88c 2.70 1)表中数据为平均数±标准误，相同菌株相同时间同列数据后不同小写字母表示差异显著（P<0.05，LSD法）
1)Datas in the table are means±SE, different lowercase letters in the same column of the same strain at the same time indicate significant difference (P<0.05, LSD method)
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表 2 真菌对禾谷孢囊线虫病室内防治效果测定
Table 2 Control efficiencies of the fungi on cereal cyst nematode in indoor pots
菌株
Strain处理1)
Treatment根长2)/cm
Root length根长增长率/%
Increased percentage of root length株高2)/cm
Plant height株高增长率/%
Increased percentage of plant hight孢囊减少率2)/%
Cyst reduction rateA1 1% 11.35±0.32b 2.53 34.43±1.25a 11.79 4.88±1.69c 2% 13.08±0.50ab 18.16 36.50±1.27a 18.51 11.65±0.97b 3% 14.26±0.60a 28.82 37.23±1.31a 20.38 31.71±1.69a CK 11.07±1.05b 30.80±1.14b B1 1% 15.67±0.67a 41.55 30.62±0.59b −0.58 7.25±2.55b 2% 15.40±1.81a 39.11 35.39±0.34a 14.90 12.19±1.24b 3% 17.67±0.76a 59.62 37.40±1.29a 21.43 36.04±0.72a CK 11.07±1.05b 30.80±1.14b 1) 处理为生防菌菌肥与病土质量比；2) 该列数据为平均数±标准误，相同菌株同列数据后不同小写字母表示差异显著（P<0.05，LSD法）
1) The mass ratio of fungus fertilizer to soil; 2) Datas in this column are means ± SE, different lowercase letters in the same column of the same strain indicate significant difference (P<0.05, LSD method)
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